Representative confocal images of an isolated cardiomyocyte obtained by SHG microscopy and by membrane fluorescent staining with di-4-ANEPPS (ANEPPS). (A) SHG (top) and ANEPPS (bottom) images with five ROIs as indicated (colored lines) following the paths of single myofibrils. The insets show the sarcomeric striation of a single ROI (cf. yellow arrows). (B) Sarcomere striation profiles taken from a single ROI using SHG and ANEPPS images. The arrows show the positions of local maxima in the intensity profiles (note their relative shift by a half sarcomere because SHG visualizes the center of sarcomeres, while ANEPPS visualizes the areas near the sarcomeric Z-disks).